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| Status |
Public on Sep 27, 2022 |
| Title |
96_OP_Repetition_1 [SAMPLE 20] |
| Sample type |
RNA |
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| Source name |
Plants without treatment
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| Organism |
Picea glauca |
| Characteristics |
sample type: Control
tissue: Needles
breeding strategy: open pollination (OP)
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| Treatment protocol |
No treatment were applied to the seedlings.
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| Growth protocol |
On January 20, 2020 seedlings were removed from cold storage and planted into 2L pots filled with Sunshine Mix #4 (Sungro, Vilna, Alberta, Canada) and grown at the Biological Sciences greenhouse, University of Alberta, for their second growing season, between January 28 and April 28, 2020 (Day 92 of the experiment). Plants were grown under natural light supplemented by cool-white fluorescent lamps (400 µmol m-2 s-1) and provided with a 16/8 photoperiod and maximum day and night temperatures of 25°C and 18°C, respectively.
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| Extracted molecule |
total RNA |
| Extraction protocol |
Using 0.5 g of ground apical internode, total RNA was extracted using the ‘Purelink RNA Mini Kit’, ‘Plant RNA Isolation Aid’ and ‘PureLink DNAse’ (Thermo Scientific Inc, Waltham, USA), following the manufacturer’s instructions The cDNA synthesis was performed using a total of 0.5 μg RNA from each sample, with the Superscript III reverse transcriptase (Thermo Scientific Inc, Waltham, USA). The PgGA2ox1, PgGA3ox1, PgGA20ox1, PgDELLA1 and PgGID1 genes were selected based on previous studies (Kayal et al. 2011, Galeano and Thomas 2020).
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| Label |
SYBR Green
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| Label protocol |
PCR reactions were performed in 10 mL, containing SYBR Green master mix [0.2 mM dNTPs, 0.3 U Platinum Taq Polymerase (Invitrogen, Waltham, USA), 0.25X SYBR Green, and 0.1X ROX], 50 ng of cDNA and 300 nM of each primer. Three technical replicates for each reaction were analyzed on an ABI PRISM 7900HT Sequence Detection System (Applied Biosystems, Waltham, USA) where the first step was 95°C for 2 min followed by 40 cycles of 95°C for 15 s and 60 °C for 1 min. The qRT-PCR experiments were done at the Molecular Biology Service Unit, University of Alberta.
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| Hybridization protocol |
n/a
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| Scan protocol |
n/a
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| Data processing |
The normalization and all the data analysis were performed according to the manufacturers instructions. For the normalization it uses the average of the TIF housekeeping gene. The three technical replicates ( PgGA2ox1, PgGA3ox1, PgGA20ox1) and two technical replicates (PgDELLA1 and PgGID1) were averaged for each sample, for the subsequent analysis. Target gene signals normalized to housekeeping gene; 2^-deltaCt, where deltaCt = (Ct_Target − Ct_HKG)].
Matrix normalized worksheet reports normalized signal (against TIF housekeeping gene).
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| Submission date |
Sep 26, 2022 |
| Last update date |
Sep 27, 2022 |
| Contact name |
Esteban Galeano |
| E-mail(s) |
esteban.galeano@msstate.edu
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| Phone |
6623257782
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| Organization name |
Mississippi State University
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| Street address |
775 Stone Blvd, 351 Thompson Hall, Mississippi State
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| City |
Starkville |
| State/province |
Mississippi |
| ZIP/Postal code |
39762-9681 |
| Country |
USA |
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| Platform ID |
GPL32694 |
| Series (1) |
| GSE214171 |
Real-time quantitative PCR analysis of white spruce (Picea glauca) families with different growth rates |
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| Supplementary data files not provided |
| Processed data are available on Series record |
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